Abstract
Assay of enzymes involved in cytokinin metabolism can be rapidly and conveniently carried out by using reversed-phase high-performance liquid chromatography (HPLC) to quantitate either the formation of product or the loss of substrate. The initial rates of loss of added 6-(3-methylbut-2-enylamino)purine or 6-benzylaminopurine can be determined in a variety of plant tissues. Asay of nucleosidase activity with 6-(3-methylbut-2-enylamino)-9-β- d-ribofuranosylpurine, 6-(4-hydroxy-3-methylbut-2-enylamino)-9-β- d-ribofuranosylpurine or adenosine as substrates can be accomplished by quantitating the loss of riboside or the formation of 8-OH cytokinins can be readily accomplished and the subsequent formation of 2,8-diOH cytokinin can be measured simultaneously if dual-wavelength monitoring is used. HPLC with a variety of solvent systems facilitates the assay of very small quantities of enzymes and the detection of multiple products.
Published Version
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